Compositions of polysaccharides derived from heparin, their preparation and pharmaceutical compositions containing them

Abstract

Alkali and alkali-earth metal salts of polysaccharides derived from heparin, their method of preparation and the pharmaceutical compositions containing them.

1. A composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin, said salts of at least one sulphated polysaccharide of heparin comprising:
 a mean molecular weight in the range of 1500 to 3000 daltons;  
 an anti-Xa activity in the range of 110 to 160 IU/mg;  
 an anti-IIa activity in the range of up to 10 IU/mg; and  
 an anti-Xa activity/anti-IIa activity ratio greater than 10:1.  
 
     
     
       2. A composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin, in which said salts of at least one sulphated polysaccharide of heparin have 2 to 26 saccharide units, have an anti-Xa activity in the range of 110 to 150 IU/mg, have a mean molecular weight in the range of 1500 to 3000 daltons, and have a 4,5-unsaturated glucuronic acid 2-O-sulphate unit on at least one end. 
     
     
       3. A composition according to  claim 1  having a mean molecular weight in the range of 2000 to 3000 daltons. 
     
     
       4. A composition according to  claim 1  having anti-Xa activity in the range of 140 to 150 IU/mg and a mean molecular weight in the range of 2000 to 3000 daltons. 
     
     
       5. A composition according to  claim 1 , in which said salts are chosen from sodium, potassium, calcium, and magnesium salts. 
     
     
       6. A composition according to  claim 1 , having an anti-IIa activity up to 5 IU/mg. 
     
     
       7. A composition according to  claim 1 , having an anti-Xa activity:anti-IIa activity ratio greater than 25. 
     
     
       8. A method of preparing at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with at least one phosphazene base;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       9. The method according to  claim 8 , in which said salts of at least one sulphated polysaccharide of heparin have a mean molecular weight in the range of 1500 to 3000 daltons. 
     
     
       10. The method according to  claim 8 , in which said salts of at least one sulphated polysaccharide of heparin have an anti-Xa activity in the range of 94 to 150 IU/mg. 
     
     
       11. The method according to  claim 8 , in which said salts of at least one sulphated polysaccharide of heparin have an anti-Xa activity up to 10 IU/mg. 
     
     
       12. The method according to  claim 8 , in which said salts of at least one sulphated polysaccharide of heparin have an anti-Xa activity:anti-IIa activity ratio greater than 10:1. 
     
     
       13. The method according to  claim 8 , in which said salts of at least one sulphated polysaccharide of heparin have 2 to 26 saccharide units and have a 4,5-unsaturated glucuronic acid 2-O-sulphate unit on at least one end. 
     
     
       14. The method according to  claim 8 , in which the quarternary ammonium salt of the benzyl ester of heparin is chosen from benzethonium, cetylpyridinium, and cetyltrimethylammonium salts. 
     
     
       15. The method according to  claim 8 , in which the at least one phosphazene base is chosen from: 
                 
 
       where R 1  to R 7  are identical or different, and are each chosen from C 1 -C 6  alkyl. 
     
     
       16. The method according to  claim 8 , in which the mol ratio of the at least one phosphazene base to the at least one quarternary ammonium salt of the benzyl ester of heparin ranges from 0.2:1 to 5:1. 
     
     
       17. The method according to  claim 8 , in which said at least one quarternary ammonium salt of the benzyl ester of heparin has a degree of esterification ranging from 50 to 100%. 
     
     
       18. The method according to  claim 8 , in which the at least one quarternary ammonium salt of the benzyl ester of depolymerized heparin is converted to a sodium salt by treating the reaction medium with an alcoholic solution of sodium acetate. 
     
     
       19. The method according to  claim 8 , in which the saponification is carried out by an alkali metal hydroxide. 
     
     
       20. The method according to  claim 8 , which the purification is carried out by hydrogen peroxide. 
     
     
       21. A method of preparing at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with sodium imidazolate;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       22. The method according to  claim 21 , in which said salts of at least one sulphated poiysaccharide of heparin have a mean molecular weight in the range from 1500 to 3000 daltons. 
     
     
       23. The method according to  claim 21 , in which said salts of at least one sulphated poiysaccharide of heparin have an anti-Xa activity in the range from 94 to 150 IU/mg. 
     
     
       24. The method according to  claim 21 , in which said salts of at least one sulphated polysaccharide of heparin have an anti-IIa activity up to 10 IU/mg. 
     
     
       25. The method according to  claim 21 , in which said salts of at least one sulphated polysaccharide of heparin have an anti-Xa activity:anti-IIa activity ratio greater than 10:1. 
     
     
       26. The method according to  claim 21 , in which said salts of at least one sulphated polysaccharide of heparin have 2 to 26 saccharide units and have a 4,5-unsaturated glucuronic acid 2-O-sulphate unit on at least one end. 
     
     
       27. The method according to  claim 21 , in which the at least one quarternary ammonium salt of the benzyl ester of heparin is chosen from benzethonium, cetylpyridinium, and cetyltrimethylammonium salts. 
     
     
       28. The method according to  claim 21 , in which the mol ratio of the sodium imidazolete to the at least one quarternary ammonium salt of the benzyl ester of heparin ranges from 0.2:1 to 5:1. 
     
     
       29. The method according to  claim 21 , in which said at least one quarternary ammonium salt of the benzyl ester of heparin has a degree of esterification ranging from 50 to 100%. 
     
     
       30. The method according to  claim 21 , in which the at least one quarternary ammonium salt of the benzyl ester of depolymerized heparin is converted to a sodium salt by treating the reaction medium with an alcoholic solution of sodium acetate. 
     
     
       31. The method according to  claim 21 , in which the saponification is carried out by an alkali metal hydroxide. 
     
     
       32. The method according to  claim 21 , in which the purification is carried out by hydrogen peroxide. 
     
     
       33. A method of treating venous thrombosis in a patient in need of such treatment, comprising administering to the patient a composition as claimed in  claim 2 , in an amount efficacious for the treatment of venous thrombosis. 
     
     
       34. A method of treating venous thrombosis in a patient in need of such treatment, comprising administering to the patient a composition as claimed in  claim 1 , in an amount efficacious for the treatment of venous thrombosis. 
     
     
       35. A method of treating arterial thrombosis in a patient in need of such treatment, comprising administering to the patient a composition as claimed in  claim 2 , in an amount efficacious for the treatment of arterial thrombosis. 
     
     
       36. A method of treating arterial thrombosis in a patient in need of such treatment, comprising administering to the patient a composition as claimed in  claim 1 , in an amount efficacious for the treatment of arterial thrombosis. 
     
     
       37. A method of treating arterial or venous thrombosis in a patient in need of such treatment comprising the administration of a solution of a pharmaceutical composition by the subcutaneous, intramuscular, intravenous, or pulmonary route, in which a composition according to  claim 2  is an active ingredient present in an amount efficacious for such treatment. 
     
     
       38. A method of treating arterial or venous thrombosis in a patient in need of such treatment comprising the administration of a solution of a pharmaceutical composition by the subcutaneous, intramuscular, intravenous, or pulmonary route, in which a composition according to  claim 1  is an active ingredient present in an amount efficacious for such treatment. 
     
     
       39. A composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin, wherein said salt is prepared according to the method of  claim 8 . 
     
     
       40. A composition according to  claim 2  having anti-Xa activity in the range of 125-150 IU/mg. 
     
     
       41. A method of treating venous thrombosis in a patient in need of such treatment, comprising administering to the patient a composition as claimed in  claim 40 , in an amount efficacious for the treatment of venous thrombosis. 
     
     
       42. A method of treating arterial thrombosis in a patient in need of such treatment, comprising administering to the patient a composition as claimed in  claim 40 , in an amount efficacious for the treatment of artarial thrombosis. 
     
     
       43. A method of treating arterial or venous thrombosis in a patient in need of such treatment comprising the administration of a solution of a pharmaceutical composition by the subcutaneous, intramuscular, intravenous, or pulmonary route, in which a composition according to  claim 40  is an active ingredient present in an amount efficacious for such treatment. 
     
     
       44. A composition according to  claim 1 , in which said at least one salt is sodium salt. 
     
     
       45. A method of preparing at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin comprising:
 depolymenzing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with 1,5,7 triazabicyclo-[4,4,0]-dec-5-ene;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       46. The method according to  claim 8 , wherein said at least one phosphazene base is 2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro-1,3,2-diaza-phosphorine. 
     
     
       47. A method of preparing at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with at least one guanidine base;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       48. A method of preparing a composition comprising the sodium salt of at least one sulphated polysaccharide of heparin, said sodium salt of at least one sulphated polysaccharide of heparin comprising:
 a mean molecular weight in the range of 1500 to 3000 daltons;  
 an anti-Xa activity in tie range of 94 to 150 IU/mg;  
 an anti-IIa is activity in the range of up to 10 IU/mg; and  
 an anti-Xa activity/anti-IIa activity ratio greater than 10:1 wherein paid method comprises:  
 depolymerizing a quarternary ammonium salt of the benzyl ester of heparin in an organic medium with a phosphazene base;  
 converting the quarternary ammonium salt of the benzyl ester of the depolymerized heparin to a sodium salt of at least one sulphated polysaccharide of heparin;  
 saponifying the sodium salt; and  
 optionally purifying the sodium salt.  
 
     
     
       49. The method according to  claim 48 , in which said sodium salt of at least one sulphated polysaccharide of heparin have an anti-Xa activity in the range of 110 to 150 UI/mg. 
     
     
       50. A method of preparing a composition comprising the sodium salt of at least one sulphated polysaccharide of heparin, said sodium salt of at least one sulphated polysaccharide of heparin comprising:
 a mean molecular weight in the range of 1500 to 3000 daltons;  
 an anti-Xa activity in the range of 94 to 150 IU/mg;  
 an anti-IIa activity in the range of up to 10 IU/mg; and  
 an anti-Xa activity/anti-IIa activity ratio greater than 10:1 wherein said method comprises:  
 depolymerizing a quarternary ammonium salt of the benzyl ester of heparin in an organic medium with 2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro-1,3,2-diaza-phosphorine;  
 converting the quarternary ammonium salt of the benzyl ester of the depolymerized heparin to a sodium salt of at least one sulphated polysaccharide of heparin;  
 saponifying the sodium salt; and  
 optionally purifying the sodium salt.  
 
     
     
       51. A method of preparing a composition comprising the sodium salt of at least one sulphated polysaccharide of heparin, said sodium salt of at least one sulphated polysaccharide of heparin comprising:
 a mean molecular weight in the range of 1500 to 3000 daltons;  
 an anti-Xa activity in the range of 94 to 150 IU/mg;  
 an anti-IIa activity in the range of up to 10 IU/mg; and  
 an anti-Xa activity/anti-IIa activity ratio greater than 10:1 wherein said method comprises:  
 depolymerizing a quarternary ammonium salt of the benzyl ester of heparin in an organic medium with 1,5,7 triazabicyclo-[4.4.0]-dec-5-ene;  
 converting the quarternary ammonium salt of the benzyl ester of the depolymerized heparin to a sodium salt of at least one sulphated polysaccharide of heparin;  
 saponifying the sodium salt; and  
 optionally purifying the sodium salt.  
 
     
     
       52. A method of preparing at least one salt chosen from alkali and alkaline-earth metal salts of sulphated polysaccharides of heparin comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with at least one base with a pKa greater than 20;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of sulphated polysaccharides of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       53. A method of preparing a sodium salt of sulphated polysaccharides of heparin comprising:
 depolymerizing a quarternary ammonium salt of the benzyl ester of heparin in an organic medium with at least one base with a pKa greater than 20;  
 converting the quarternary ammonium salt of the benzyl ester of the depolymerized heparin to the sodium salt of sulphated polysaccharides of heparin;  
 saponifying the sodium salt; and  
 optionally purifying the sodium salt.  
 
     
     
       54. The method according to  claim 53 , wherein said depolymerizing step is accomplished with a single base with a pKa greater than 20. 
     
     
       55. The method according to  claim 54 , wherein said base is 2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro-1,3,2-diaza-phosphorine. 
     
     
       56. The method according to  claim 8 , in which the at least one phosphazene base is chosen from: 
                 
 
       where R 1  to R 7  are identical or different, and are each chosen from C 1 -C 6  alkyl and, further where R 3  and R 4  or R 1  and R 7 , taken together with the nitrogens to which they are attached, may form a saturated ring chosen from substituted and unsubstituted six member rings. 
     
     
       57. The method according to  claim 47 , in which the at least one base of guanidine comprises: 
                 
 
       where R 1  is chosen from hydrogen and alkyl, and where R 2 , R 3 , R 4 , and R 5 , which are identical or different, are each chosen from C 1 -C 6  alkyl. 
     
     
       58. The method according to  claim 57 , where R 1  is hydrogen, and R 2 , R 3 , R 4 , and R 5  are each methyl. 
     
     
       59. A composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin, said salts of at least one sulphated polysaccharide of heparin comprising:
 a mean molecular weight in the range of 1500 to 3000 daltons;  
 an anti-Xa activity in the range of 110 to 150 IU/mg;  
 an anti-IIa activity in the range of up to 10 IU/mg; and  
 an anti-Xa activity/anti-IIa activity ratio greater than 10:1;  
 
       wherein said salts of at least one sulphated polysaccharide of heparin have a 4,5-unsaturated glucuronic acid 2-O-sulphate unit on at least one end. 
     
     
       60. A method of treating venous thrombosis in a patient in need of such treatment, comprising administering to the patient a composition as claimed in  claim 59 , in an amount efficacious for the treatment of venous thrombosis. 
     
     
       61. A method of treating arterial thrombosis in a patient in need of such treatment, comprising administering to the patient a composition as claimed in  claim 59 , in an amount efficacious for the treatment of arterial thrombosis. 
     
     
       62. A method of treating arterial or venous thrombosis in a patient in need of such treatment comprising the administration of a solution of a pharmaceutical composition by the subcutaneous, intramuscular, intravenous, or pulmonary route, in which a composition according to  claim 59  is an active ingredient present in an amount efficacious for such treatment. 
     
     
       63. A method of preparing a composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin as claimed in  claim 1 , the method comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with at least one phosphazene base;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       64. A method of preparing a composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin as claimed in  claim 1 , the method comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with sodium imidazolate;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       65. A method of preparing a composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin as claimed in  claim 1 , the method comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with 1,5,7 triazabicyclo-[4.4.0]-dec-5-ene;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       66. A method of preparing a composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin as claimed in  claim 1 , the method comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with at least one guanidine base;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharlde of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.  
 
     
     
       67. A method of preparing a composition comprising at least one salt chosen from alkali and alkaline-earth metal salts of at least one sulphated polysaccharide of heparin as claimed in  claim 1 , the method comprising:
 depolymerizing at least one quarternary ammonium salt of the benzyl ester of heparin in an organic medium with at least one base with a pKa greater than 20;  
 converting the at least one quarternary ammonium salt of the benzyl ester of the depolymerized heparin to at least one salt chosen from alkali and alkaline-earth metal salts of sulphated polysaccharides of heparin;  
 saponifying the at least one salt; and  
 optionally purifying the at least one salt.